Modified starch material of biocompatible hemostasis

ABSTRACT

A modified starch material is arranged for biocompatible hemostasis, biocompatible adhesion prevention, tissue healing promotion, absorbable surgical wound sealing and tissue bonding, when applied as a biocompatible modified starch to the tissue of animals. The modified starch material produces hemostasis, reduces bleeding of the wound, extravasation of blood and tissue exudation, preserves the wound surface or the wound in relative wetness or dryness, inhibits the growth of bacteria and inflammatory response, minimizes tissue inflammation, and relieves patient pain. Any excess modified starch not involved in hemostatic activity is readily dissolved and rinsed away through saline irrigation during operation. After treatment of surgical wounds, combat wounds, trauma and emergency wounds, the modified starch hemostatic material is rapidly absorbed by the body without the complications associated with gauze and bandage removal.

CROSS REFERENCE OF RELATED APPLICATION

This is a Divisional application that claims the benefit of priorityunder 35 U.S.C. §119 to a non-provisional application, application Ser.No. 12/228,029, filed Aug. 8, 2008.

BACKGROUND OF THE INVENTION

Field of Invention

The present invention relates to a modified starch material ofbiocompatible hemostasis, biocompatible adhesion prevention, tissuehealing promotion, absorbable surgical sealing and tissue bonding, andmore particularly to a modified starch material, which is absorbable byhuman beings and animals, and applied directly to wound surface ofhumans and mammals, including wound surface with blood or extravasate,to stanch blood, prevent adhesion, promote tissue healing, seal sectionof wound tissue, prevent bleeding and exudation of tissue fluid, bondtissue and organ wounded by trauma or operation, help repairing tissue,and avoid or reduce surgical suture.

Description of Related Arts

Surgical operations and trauma may create bleeding wounds, which canproduce a risk of excess blood loss. Therefore, hemostats to controlbleeding should be applied in a timely manner. It is a common to applybiocompatible, absorbable hemostatic agents to bleeding wound sites toachieve hemostasis (cessation of bleeding) in surgical procedures,trauma treatment and home self rescue. There is clinical benefit toprovide patients a hemostatic agent which is safe, efficacious, easy touse, and cost effective.

Prior absorbable hemostats consist of the following classes ofmaterials:

Hemostatic sponge class: gelatin sponge, collagen sponge, chitosansponge, carboxymethyl cellulose sponge, thrombin and fibrin sponges;

Hemostatic gauze/hemostatic film class: oxidized cellulose gauze,oxidized regenerated cellulose gauze, oxidized cellulose gauze withcarboxymethyl cellulose;

Hemostatic glue class: fibrin glue, synthetic glue;

Polysaccharide hemostatic powder class: microporous polysaccharidepowder, chitosan powder, algae powder.

A detailed analysis of absorbable hemostats in common use is statedbelow:

1. Absorbable Gelatin Sponges and Collagen Sponges:

The gelatin sponge is extracted from animal tissue, and the maincomponent of the gelatin sponge is animal collagen. The gelatin spongehas a hydrophilic and multi-porous structure to concentrate bloodcomponents by absorbing water in the blood to arrest bleeding. However,gelatin is a collagen-based material from animal extract and containsheterogenetic protein which may cause anaphylaxis, resulting in feverishsymptoms in patients. Further, the human body absorbs the gelatinmaterial very slowly, and on average requires more than four weeks tofully dissolve. Foreign agents with slow absorption times can be sitesfor infection, tissue inflammation, and wound healing retardation.

Collagen sponges, which are also extracted from animal tissue, promoteblood coagulation by activating the endogenous coagulation cascade whilealso concentrating blood components by absorbing water in the blood.

Like the gelatin sponges, collagen sponges are also sourced from animalcollagen and contain heterogenetic protein, which is slow to absorb inthe human body. The collagen sponge may produce complications ofanaphylaxis, slow healing and infections. Due to these clinical risks,applications of collagen sponges may be limited in the future.

2. Oxidized Cellulose Hemostatic Gauze and Oxidized RegeneratedCellulose Hemostatic Gauze:

Oxidized cellulose is a cellulose derivative. The hemostatic mechanismof oxidized cellulose is the concentration of blood components throughthe hygroscopic activity of oxidized cellulose, which stimulates bloodcoagulation as the carboxyl material combines with haemoglobin Fe toproduce acidic hematin in the blood. The resulting brown gel sealscapillary vessels and promotes hemostasis. Oxidized regeneratedcellulose has the same mode of action as oxidized cellulose.

Oxidized cellulose is synthetic. Normal human tissue degrades oxidizedcellulose slowly by metabolizing enzymes. This process generallyrequires 3-6 weeks depending on the dosage and the tissue location inthe body. Oxidized cellulose may cause local infection and adverselyaffect local tissue healing. Patent application, China publicationnumber CN1533751A, discloses a hemostatic wound dressing with a tradename of SURGICEL. SURGICEL includes a cellulose fabric and amulti-porous polymer substrate on the fabric surface which contacts thewound. The substrate contains biocompatible, water-soluble polymers. Thefabric fibers are oxidized regenerated cellulose and the biocompatible,water-soluble polymers are polysaccharides. This hemostatic wounddressing consists primarily of oxidized cellulose, a slowly absorbingmaterial in the human body.

3. Fibrin Glues:

Fibrin glues consist of fibrinogen, thrombin, aprotinin and calciumchloride. The hemostatic action relies mainly on the activation offibrinogen by the thrombin to promote coagulation cascade. Fibrinsealants are a mixture of fibrinogen and thrombin and have been widelyused in recent years. The thrombin and fibrin in fibrin glues aresourced from either animal or human blood components and thereforecreate the risk of anaphylaxis and viral infections such as hepatitis,AIDS, and BSE. Fibrin glues demonstrate weak adhesion when applied towet, bleeding tissue and may be ineffective in the presence of activebleeding. Further, fibrin glues require special mixing, timing andstorage condition.

4. Natural Biological Polysaccharide Products:

In recent years, natural, biological polysaccharide-based products havefocused much attention. The natural biological polysaccharide productsare derived from plant material and chitosans and usually presented inpowder form. These products have good biocompatibility, no toxicity, notissue irritation, and no risk of anaphylaxis or viral infection fromanimal or human components contained in other hemostats.

Chitosan/Chitin Products:

Chitosan products are typically available in high swelling andnon-absorbable sponges. Chitosan is made from the crushed shells ofcrustaceans. Chitosan has rapid hydrophilic capability and can activatethe blood coagulation mechanism through its strong ionic charge.However, due to a lack of human enzymes to degrade chitosan,chitosan-derived products will be confined to topical applications.There is no evidence that chitosan products have been used in clinic asabsorbable surgical hemostats.

Microporous Polysaccharide Hemospheres (MPH):

In 2002, MEDAFOR, INC. in the USA developed an absorbable hemostaticmaterial called Arista™ (U.S. Pat. No. 6,060,461), which consists ofmicroporous polysaccharid particles (MPH). The microporouspolysaccharide particles are made through the reaction of purifiedstarch and epichlorohydrin, wherein the epichlorohydrin reacts withstarch molecules. This reaction results in the formation of ethylpropanetriol which creates a glucose molecule crosslink to the 3Dnetwork structure.

There are a few disadvantages of the MPH hemostatic powder. Firstly, thedelivery of MPH mainly focuses on local, easy-to-access wound sites butpresents some difficulties for effective applications for deep, tortuouswounds, in particular the endoscopic procedures (such as minimallyinvasive surgery via endoscope and laparoscope). Secondly, during theproduction process, epichlorohydrin, a colorless, oily and toxicchemical, is employed to produce a required reaction. This productionprocess is not environment friendly. The cost of production isrelatively expensive. Thirdly, the hemostatic efficacy of MPH is notsatisfactory in particular for profuse bleeding due to its lowhydrophilic capacity and slow water absorption characteristic. Fourthly,the adhesiveness of the MPH to tissue is low following contacting withblood. The low viscosity, low adhesiveness of MPH following waterabsorption may reduce the hemostatic efficacy of MPH due to its weaksealing capability to wounded tissues and broken vessels. Fifthly, inthe presence of active bleeding, the MPH powder can be easily washedaway by blood flow if not compressed with a gauze on the top of powder.This gauze compression requirement adds an additional step in thehemostatic powder application technique and may risk re-bleeding whenthe gauze is removed. Therefore, MPH may have an unsatisfactoryhemostatic efficacy for active bleeding.

SUMMARY OF THE INVENTION

An object of the present invention is to provide a biocompatible,modified starch composition and new uses of the modified starch inhumans and/or animals as a topical and surgical hemostat. Hemostasisoccurs immediately and effectively when the said hemostat contacts bloodon wound sites.

Another object of the present invention is to provide a modified starchcomposition as an agent for anti-adhesion therapy, for promoting tissuehealing, for sealing wounds and bleeding vessels, for adhesive sealingand tissue bonding, and for promoting bacteriostatic andanti-inflammatory effects on bleeding wound sites. In addition to itshemostatic performance, the application of the invention will supportanti-adhesions, tissue healing, surgical sealing and tissue bonding andreinforcement when the bleeding wound is on the skin surface or in theinternal organs and whether the application is in an open surgicaloperation, trauma treatment, or delivered under laryngoscope, endoscope,and laparoscope.

Another object of the present invention is to provide methods ofproducing the modified starch hemostatic composition and thetechnological formula to manufacture the modified starch in thefollowing formats: powder, sponge, foam, gel, film and others. Theseformats do fulfill dynamic surgical hemostatic requirements, and areeasy to use.

Another object of the present invention is to provide the methods,process and techniques to modify the starch composition to satisfyimportant physical and chemical properties and characteristics,technical parameters and technical indices required for a hemostat,surgical sealant, an agent for anti-adhesion, tissue healing, tissueadhesive sealing and tissue bonding. The modified starch hemostaticcomposition is absorbed by humans and animals, is safe and effective,and can be degraded rapidly.

In addition, the modified starch in the present invention can be used asa biocompatible, an anti-adhesion material, a tissue healing agent, asurgical sealant, and a tissue bonding/reinforcement substance fortissue repair.

The technical formulas in the present invention fulfill the foregoingperformance requirements with modified starch applications as abiocompatible hemostatic material with mechanisms that includedissolving or swelling in water and the subsequent formation of adhesiveglue or adhesive gel.

The mechanism further includes a modified starch acquisition ofhydrophilic groups in its molecular chains through the modificationprocess.

When the hydrophilic and enhanced adhesive modified starch according tothe present invention is applied to bleeding wound sites, it rapidlyabsorbs water in the blood and concentrates blood components. Meanwhile,this interaction creates an adhesive matrix formed with the blood andplasma which adheres to the bleeding wound, mechanically seals thebroken blood vessels and stops bleeding.

The modified starch material according to the present invention includesstarch modified physically, chemically, naturally, or enzymatically, andstarch modified repeatedly with at least one of the above methods or acombination of two or more of the above methods.

The physical modifying process according to the present inventioncomprises irradiation, mechanical, and steam treatment.

Physically modified starch, for example, a pre-gelatinized starchtreated solely with spray drying or irradiation process, is remarkablysafe as a bio-absorbable, hemostatic material since it is not treatedwith any chemical agents.

The starch can be pre-gelatinized by the following physical modifyingprocesses: a) dry-process, such as an extrusion process and a rollerprocess; b) wet-process, such as a spray drying process.

Specifically, after heating the raw starch with a measured amount ofwater, starch granules swell to a pasty substance, regularly arrangedmicelle of starch are broken, crystallites disappear, and the resultingcomposition is easily degraded under the process of amylase. Thepre-gelatinized starch is able to swell and/or dissolve in cold or roomtemperature water and form an adhesive paste whose retrogradation islower than that of raw starch, affording easier handling during theproduction process.

Raw starch can be pre-gelatinized through solely a physical modificationprocess without adding any chemical agents and becomes a hemostaticmaterial with enhanced hydrophilic and adhesive properties.

The pre-gelatinized starch of the present invention is safe, non-toxic,and has no adverse side effects. The pre-gelatinized starch is readilydegraded and metabolized by enzymes in the body. The pre-gelatinizedmaterial of the present invention is safe and biocompatible.

The chemical modifying according to the present invention includesacidolysis, oxidation, esterification, etherification, cross-linking,chemical agent grafting, or multiple modifying processes including atleast two of the above processes, or one of the above modifyingprocesses performed at least twice.

In the present invention, by adding the functional group on the rawstarch glucose units with chemical agents, e.g. by carboxylationmodification, or hydroxylation modification, the starch captureshydrophilic groups in its molecular structure and obtains hydrophilicproperties. By using bifunctional or polyfunctional chemical agents tocross-link the raw starch macromolecules or grafting externalmacromolecurlar hydrophilic groups to the raw starch, the starchacquires enhanced hydrophilic properties and viscosity/adhesiveness in awater solution. The viscosity of modified starch relates to the rawstarch origin and the degree of substitution of external andcross-linked or grafted functional groups, etc. . . . . When contactingblood, the hydrophilic and adhesive properties of the modified starch ofthe present invention produce a “starch-blood coagulation matrix” withstrong adhesive characteristics which can seal wounded tissue and stopbleeding. In addition, the interaction between the formed bloodcoagulation matrix and the functional groups of tissue proteins causesthe “starch-blood coagulation matrix” to adhere to and seal the woundedtissue, resulting in hemostasis.

Specifically, the described modified starch contains one or more groupsof pre-gelatinized starch, acid modified starch, dextrin, oxidizedstarch, esterified starch, etherified starch, and cross-linked starch.

The described hemostatic composition comprises two or more modifiedstarches to satisfy the physical and chemical properties of a hemostaticagent, where the weight ratio of the two modified starch groups can be99:1˜1:99.

Specifically, the weight ratio of the two modified starch groups can be:95:5, 90:10, 85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, or 50:50.

The main physical parameters of the modified starch, according to thepresent invention, are provided below:

The modified starch of the present invention has a molecular weight over15,000 Daltons (for instance, 15,000˜2,000,000 Daltons).

The modified starch of the present invention has a water absorbencycapacity not lower than one time its weight (i.e. 1 gram of describedmodified starch can absorb 1 gram or more of water); whereas it can be1˜500 times generally and 2˜100 times preferably.

The modified starch composition of the present invention includes, butnot limited to, at least one carboxymethyl starch, hydroxyethyl starch,and cationic starch.

For example, the carboxymethyl starch is a polymer of linear structureas expressed in the following formula:

Modified starches such as carboxymethyl starch (CMS) and hydroxyethylstarch are known clinically as plasma substitutes. These modifiedstarches exhibit biocompatibility and safety with no toxic side effectswhen employed in the human circulatory system. The hemostaticcomposition, according to the present invention, can further includeother plasma substitutes by means of well-known pharmacokineticsapproaches and specified physical/chemical properties to produce safeand reliable hemostatic agents.

When cationic starch of the modified starches is selected as ahemostatic material, the surface positive charge of the cationic starchattracts and interacts with electronegative blood erythrocytes,accelerating the blood coagulation process. Furthermore, when contactingblood, the positively charged modified starch adheres tightly to tissue,seals the wound, and rapidly stops bleeding. The cationic starch can beused independently as a hemostatic material or mixed with other modifiedstarches as a composite hemostatic material.

Composite modified starch comprises, but is not limited to, at leastpre-gelatinized hydroxypropyl distarch phosphate. Specifically, thehydroxypropyl distarch phosphate is produced by cross-linking andetherifying the starch with propylene oxide and phosphoric acid,followed by pre-gelatinization modification through a spray dryingprocess. The hydroxypropyl distarch phosphate of the present inventionhas high adhesiveness, strong water absorbency and robust hemostaticeffects. It is stable in acidic or alkali environments and can be usedas a biocompatible, hemostatic material, a surgical sealant, a tissuehealing composition, an anti-adhesion agent, and a tissue repairmaterial.

The cross-linked starch of the present invention includes, but is notlimited to, at least one of epichlorohydrin cross-linked starch andcross-linked carboxymethyl starch.

The grafted starches of the present invention include at least apropylene ester-carboxymethyl starch grafted copolymer and a crylicacid-carboxymethyl starch grafted copolymer. Grafted starch has bothenhanced water absorption capability and high viscosity/adhesiveness.Therefore, it has a profound effect on hemostasis when applied to woundsurfaces, especially combat wounds, traumatic wounds, and profusebleeding from large arteries and large veins due to aneurysms or largephlebangioma ruptures.

The modified starch, according to the present invention, can be made inpowder form, spherical form or aerosol form to be delivered directly tothe bleeding wound surface.

As to the wound surface of large burn areas, adopting inhalator oraerosol can stanch blood at the wound surface, reduce tissue fluidexudation, and keep the wound surface moist to aid tissue healing.

In addition, the hemostatic material of the present invention can bemade into hemostatic sponge, foam, film, and plaster, which can beapplied to a bleeding wound site to stanch blood directly, wherein thehemostatic sponge, foam, the hemostatic film, and the hemostatic plastercan be made into a film or an attaching layer to the inside or surfaceof a fiber fabric, such as a bandage, band-aid; etc. Such hemostaticsponge, foam, hemostatic film, and hemostatic plaster can be columnar,sheet, massive, flocculent, or membranous.

The modified starch hemostatic foam of the present invention is easy toapply to active bleeding sites and achieves an optimal hemostaticoutcome. The hemostatic foam of the present invention can be made fromone or more varieties of modified starch processed by vacuum freezedrying. The hemostatic foam of the present invention can be compositehemostatic foam made from one or more varieties of modified starches andother biocompatible hemostatic materials processed by vacuum freezedrying or other drying processes.

Wherein, according to the present invention, other biocompatiblehemostatic materials other than the modified starches can comprise oneor more of the groups of gelatin, collagen, carboxymethyl cellulose,oxidized cellulose, oxidized regenerated cellulose, and chitosan.

In order to solve the challenge of molding modified starch into spongesand foam, the present invention combines other known bioabsorbablehemostatic materials with strong biocompatibility and clinicallyacceptable qualities with the modified starch of the present inventionto produce composite hemostatic sponges and foams. Whereas other knownbioabsorbable hemostatic materials can be of one or more components, themodified starches can also be of one or more components, such asmodified starch+gelatin, modified starch+collagen, modifiedstarch+thrombin, modified starch+chitosan, modified starch+carboxymethylcellulose, and modified starch+hyaluronic acid. These combinations canbe molded into sponge and foam forms to satisfy clinical requirements.

Weight proportions between the biocompatible modified starch and otherbiocompatible hemostatic materials can be 99.9:0.1˜1:99.

Specifically, the weight ratio between the modified starch and otherbiocompatible hemostatic materials preferably is: 95:5, 90:10, 85:15,80:20, 75:25, 70:30, 65:35, 60:40, 55:45, 50:50, 45:55, 40:60, 35:65,30:70, 25:75, or 20:80.

This additional coagulant material may be added to the describedmodified starch hemostatic sponge or foam directly during the vacuumfreeze drying production process to produce a composite hemostaticsponge or composite hemostatic foam. The production process may involve,but is not limited to, pre-mixing the coagulant material with themodified starch directly before vacuum freeze drying process.

Accordingly, the coagulant of the present invention comprises one ormore combinations of the following group of blood coagulation factors:thrombin, fibrin, calcium agent, polypeptide, peptide, amino acid, andprotamine.

The modified starch sponge and foam of the present invention can bemanufactured into hemostatic sponges and foam formed by a vacuum freezedrying process utilizing a forming agent or a plasticizing agent.

Whereas the forming agents of the present invention comprises, but notlimited to, organic forming agents, inorganic forming agents, naturalforming agents, and man-made plasticizing agents, which may include, butnot limited to, one or more combinations of glycerol, kaolin, sorbitol,ethanol, ammonia, and polyethylene glycol.

Specifically, the vacuum freeze drying process is a drying method thatfreezes wet material or solutions to a solid state under lowtemperatures (−10˜−50° C.) and then converts the solid material into agas and then, in a vacuum (1.3-1.5 Pa), back to a solid material withoutan intermediate liquid phase (sublimination). As the vacuum freezedrying is processed under low temperature and low pressure, the moisturesublimes directly to produce a substance with numerous specialproperties. The basic parameters of the vacuum freeze drying processspecify both physical parameters and process parameters. The physicalparameters include thermal conductivity, transfer coefficient, etc. Theprocess parameters include freezing, heating, state of the material,etc. Continued research on this freezing process involves experiments toidentify the optimal freezing curve. As the described biocompatible,modified starch hemostatic material can be made into a hemostatic glue,the physical form can be colloidal, dissolved colloidal, thawedcolloidal, semi-liquid or gelatinous, etc. The hemostatic glue can beproduced by adding other liquids, not limited to water, to the modifiedstarch by diluting, swelling, or dissolving the liquids in certainproportions.

According to the present invention, the topical application of modifiedstarch can be used as a hemostatic agent to manage and control bleedingwound surfaces in humans, mammals, birds, or reptiles, and the internalapplication for hemostasis of bleeding wound surfaces within humanbodies, tissues and organs following surgical operations and traumatreatments, under open surgery or nasoscope, laryngoscope, endoscope andlaparoscope.

The modified starch hemostatic composition, according to the presentinvention, can be applied for hemostasis on bleeding bone tissue causedby surgery or trauma, particularly for hemostasis in spongy bone tissue.In thoracic or neurological operations involving some patients, such aschildren, elderly people, and patients with osteoporosis, sternalbleeding and skull bleeding is difficult to control. It is common toapply bonewax to the sternum or skull, however, bonewax is slow toabsorb and may cause complications such as non-union or infection. Themodified starch composition, according to the present invention, is abiocompatible substitute for bonewax to control bone bleeding andmechanically seal the wound caused by surgery or trauma with its robusthydrophilic properties, strong adhesiveness and ease of molding. Themodified starch hemostatic material degrades rapidly after surgery andavoids the complicating issues of non-union and infection associatedwith bonewax. When the modified starch is applied as a hemostaticmaterial, other biological benefits of the described modified starch areworthy of attention. It is essential to evaluate the described modifiedstarch composition as having further positive effects on woundinflammation, tissue adhesion and tissue healing while acting as ahemostat.

It is proven that the modified starch hemostatic material, according tothe present invention, has further application as an absorbable,postoperative tissue adhesion barrier. The adhesion barrier of themodified starch, according to the present invention, prevents woundedtissue or organs from adhering to other tissue or organs in thevicinity, thereby reducing local bleeding and exudation, by mechanicallyisolating the wound or wound surface from adjacent tissue andsurrounding organs, such as the peritoneum.

The modified starch, according to the present invention, can promotetissue healing, including skin, subcutaneous soft tissue, musculature,bone tissue, neurological tissue, nerve tissue, and wounded tissue ofthe liver, kidney, spleen, etc., through proper dosage and application.The modified starch can be a “scaffold” for skin tissue cells to promotehealing and the growth of skin tissue from large wound surfaces due toburns, a “scaffold” for osteocyte growth and propagation in bone defectsfrom trauma, bone tumor resection, etc., and a “scaffold” forneurological tissue cell growth and propagation when applied to injuredneurological tissue caused by brain trauma, brain tumor resection, etc.The modified starch, according to the present invention, has furtherapplications as a biocompatible surgical sealant, capable of forming aprotective layer of colloid or film on the wound surface to seal andprevent drainage of blood, tissue fluids, lymph fluid, cerebrospinalfluid, bile, gastric fluid, and other intestinal fluids resulting fromsurgery and trauma treatment. This sealing effect will prevent lymphfistula leakage, bile flaccidity, pleural flaccidity, intestinalflaccidity, cerebrospinal flaccidity, and vascular flaccidity.

The modified starch, according to the present invention, has furtherapplication as a biocompatible tissue adhesive, capable of adhering,repairing, and bonding wounded nerve tissue, musculature, and tissues ofthe bone, skin, viscera, and subcutaneous tissue. It can also bond othercurative materials to wounded tissue and organs for tissue repair.

In addition to the above advantages, the modified starch of the presentinvention has a bacteriostatic and anti-inflammatory effect on bleedingwound surfaces. The modified starch hemostatic material, according tothe present invention, has a hemostatic effect which controls bleeding,reduces blood and tissue fluid exudation, and maintains a moist woundsurface. As a result, it suppresses the growth of bacteria and reducesthe inflammatory response, diminishing local irritation and relievingpain. Furthermore, to strengthen the anti-inflammatory response, knownantibiotic or other anti-pyrotic agents can be added to the modifiedstarch during the manufacturing process to produce hemostatic powder,sponges and foams, hemostatic glues and gels, etc., all of which aresuitable for topical and internal clinical applications.

Another advantage of the modified starch hemostatic material of thepresent invention is the rapid particle dissolution in water,facilitating the easy removal of excess modified starch particles fromthe wound by simple saline irrigation. The residual modified starch notactively involved in hemostasis can be rinsed away by irrigation. In thetreatment of battle wounds, self rescue, or first aid, the hemostaticmaterial remaining in small amounts will be absorbed by the body and theirritation of wound debridement or gauze removal is avoided.

The modified starch hemostatic material has properties of stability,extended shelf life, resistance to high and low pressure, resistance tohigh temperature (up to 60° C.) and low temperature (down to −40° C.),convenient storage, and physical stability. Therefore, it may also beemployed as a hemostatic material for the military, emergency, andfirst-aid uses. Particularly, it can be adapted for extremeenvironmental conditions such as desert areas, polar regions, alpineareas, outer space, and underwater probes.

The modified starch sponge and foam has physical properties ofpliability, flexibility, moldability, and curling. It can be adapted forwound surfaces with various shapes, sizes, and features, such as deepand irregular anatomical wounds, organ physiologies, both inside andoutside the lacuna surface, and applied under endoscope, laparoscope oropen surgery.

To enhance the safety of applying the modified starch to wound surfaces,tissue, etc., the modified starch material, according to the presentinvention, can be packaged and sterilized with, but not limited to,gamma irradiation, oxirane, and ozone sterilization.

At least a production process of a biocompatible modified starchmaterial according to the present invention, comprising the steps of:

providing a modified hygroscopic biocompatible starch material andadding into a agglomerator under 40˜50° C.; and

adding distilled water and producing a modified starch finished productmaterial by particle agglomerating and pellet processing;

wherein the modified starch finished product has a molecular weight over15,000 Daltons (for instance, 15,000˜2,000,000 Daltons) and a graindiameter of 10˜1000 μm, wherein starch grains with diameters of 30˜500μm represent no less than 95% of the total amount of starch grains,wherein the modified starch finished product can provide effects ofhemostasis, adhesion prevention, tissue healing promotion, sealing,adhesive plugging, bonding to a bleeding wound surface, andbacteriostatic and anti-inflammatory effect on the bleeding woundsurface of either external or internal bleeding tissue and organs. Themodified starch finished product can be applied for topical use, forsurgical use, or via laryngoscope, endoscope and laparoscope.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates hemostatic effects of Arista™ in a rabbit liverbleeding model.

FIG. 2 illustrates hemostatic effects of a modified starchcarboxymethyl-starch 66# in a rabbit liver bleeding model.

FIG. 3 illustrates hemostatic effects of raw starch in a rabbit liverbleeding model.

FIG. 4 illustrates adhesion in abdominal cavity in the experimentalgroup (carboxymethyl starch 66#) 24 hours after the establishment of themouse model.

FIG. 5 illustrates degradation in abdominal cavity in the experimentalgroup (carboxymethyl starch 66#) 24 hours after the establishment of themouse model.

FIG. 6 illustrates intestinal adhesion in the control rat group (blank).

FIG. 7 illustrates preventive effects of modified starch-carboxymethylstarch 66# on intestinal adhesion in rats.

FIG. 8 illustrates preventive effects of sodium hyaluronate (positivecontrol) on intestinal adhesion in rats.

FIG. 9 illustrates a comparison of bone healing indexes in rabbits.

FIG. 10 illustrates a representative scanning electron microscope photoof section of pre-gelatinized hydroxypropyl distarch phosphate (51#)hemostatic sponge A.

FIG. 11 illustrates a representative scanning electron microscope photoof section of pre-gelatinized hydroxypropyl distarch phosphate (51#)hemostatic sponge B.

FIG. 12 illustrates a comparison of water absorption ability betweencarboxymethyl starch 66# and Arista™.

FIG. 13 illustrates a comparison of work of adhesion among carboxymethylstarch 66#, hydroxyethyl starch 88#, and Arista™.

FIG. 14 illustrates a comparison of viscosity between carboxymethylstarch 66#, according to the present invention, and Arista™.

FIG. 15 illustrates water absorbency of various modified starch by themethod of centrifugation.

FIG. 16 illustrates hemostatic effects of the materials on experimentalanimals under different hemostatic conditions.

FIG. 17 illustrates grades of intestinal adhesion in different ratgroups.

FIG. 18 illustrates results of seven bone healing indexes in rabbitgroups.

FIG. 19 illustrates a comparison of physical and chemical propertiesamong the above-mentioned hemostatic sponges and other hemostaticsponges

FIG. 20 illustrates water absorption ability of composite hemostaticsponges.

FIG. 21 shows a comparison of water absorbency between compositehemostatic sponges and other hemostatic sponges.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

The present invention provides a modified starch material with thebiocompatible properties of hemostasis, adhesion prevention, tissuehealing, absorbable tissue sealing and tissue bonding. Morespecifically, the modified starch material, which is rapidly degraded byhumans and animals when applied directly to the wound surface of humansand mammals, including a bleeding or exudating wound surface, is able tostop bleeding, prevent adhesions, promote tissue healing, seal woundedtissue, prevent bleeding and fluid exudation from tissue, bond andrepair tissue or organs injured during trauma or surgery, and avoid orminimize surgical sutures.

Starch is a glucosan. At room temperature, raw starch is generally notsoluble in water, nor does it readily absorb water. Raw starch normallyabsorbs water at temperatures above 60° C. and swells to an adhesive,translucent and colloidal solution. The modified starch is raw starchprocessed through physical and chemical modifications, resulting inphysical and chemical changes which make its characteristics andproperties suitable for applications in various industries. Starch isgenerally classified by its origin, such as potato starch or cornstarch, etc. There are two types of glucose chains in starch, includinga simple chain called amylose and a complex branched form calledamylopectin. The diameter range of starch grains is normally between1˜100 μm and the average diameter is from 15 to 30 μm.

Natural raw starch has minimal hemostatic characteristics because starchgrains are small and light and the hydrophilic properties areunsatisfactory at room temperature.

The modified starch acquires certain chemical and physicalcharacteristics by cutting, rearranging, or adding other chemical groupsthat change the structure of the raw starch molecular chain. Themodified starch can be categorized primarily into physically modifiedstarch, chemically modified starch, enzymatically modified starch, andnaturally modified starch, according to the performed modificationprocess.

Physical modification is the process which produces modified starch,with the desired properties and functions, by physically changing themicrocrystalline starch structure through heating, extrusion, andirradiation. Specifically, physically modified starch includes ofpre-gelatinized starch (α-starch), γ-ray, microwave or high frequencyradiation starch, mechanically milled starch, and steam treated starch,etc.

Chemically modified starch is produced by processing the raw starch withchemical agents and changing the molecular structure to achieve thedesired modified starch properties. Specifically, the chemicallymodified starch is categorized primarily as acid modified starch,oxidized starch, baking dextrin, esterified starch, etherified starch,grafting starch, etc.

Enzymatically modified starch is produced by processing the raw starchwith enzymes, such as α-cyclic dextrin, β-cyclic dextrin, γ-cyclicdextrin, malto-dextrin and amylopectin.

Naturally modified starch may possess the same properties as chemicallymodified starch by changing the structure of natural raw starch with avariety breeding and genetic techniques.

The modified starches generally require multi-modification of the rawstarch to achieve the desired properties. In another words, it is themodification with two or more modifying methods that produces the final,composite, modified starch. Most of the widely used modified starchesare composite modified starches that have been modified several times.

The modified starch material according to the present invention can beapplied to a bleeding wound surface in humans and animals as ahemostatic agent for topical and surgical use. The modified starchmaterial according to the present invention can be used on soft tissueand organs to rapidly and effectively control bleeding.

The present invention provides methods and technological approaches ofproducing the hemostatic modified starch material, which produce themodified starch in form of powder, sponge, foam, colloid, film, andother forms that satisfy various surgical hemostatic requirements,including ease of use.

The present invention provides biocompatible modified starch materialwhich can be produced by various methods and processes to achieveessential physical and chemical properties, characteristics, technicalparameters, and technical indices required for hemostasis, adhesionprevention, sealing, adhesive gluing, promotion of healing, and tissuebonding within the application environment. The modified starchhemostatic material is safe, reliable, absorbable, and rapidlydegradable by humans and animals.

Additionally, the modified starch material of the present invention canbe used as a biocompatible anti-adhesion agent, a tissue healingpromotion material, a surgical sealant, and a tissue bonding compositionfor tissue repair.

The technical formulas of the present invention fulfill the foregoingperformance requirements with modified starch applications as abiocompatible hemostatic material with characteristics that includedissolving or swelling in water and the subsequent formation of anadhesive glue or adhesive gel.

The characteristics of the modified starch of the present inventionfurther include the acquisition of hydrophilic groups in its molecularchains through the described modification process.

When the hydrophilic and enhanced adhesive modified starch is applied tobleeding wound sites, it rapidly absorbs water in the blood andconcentrates blood components. Concurrently, this interaction creates anadhesive matrix formed with the blood and plasma which adheres to thebleeding wound, mechanically seals broken blood vessels and stopsbleeding.

The modified starch material according to the present invention includesstarch modified physically, chemically, naturally, or enzymatically, andstarch modified repeatedly with at least one of the above methods or acombination of two or more of the above methods.

The physical modifying process according to the present inventionemploys irradiation, mechanical, and steam modification.

Physically modified starch, for example, a pre-gelatinized starchtreated solely with spray drying or irradiation process, is remarkablysafe as a bio-absorbable, hemostatic material since it is not treatedwith any chemical agents.

Pre-gelatinized starch can be modified by an extrusion process, rollerdrying, and a spray drying process.

Specifically, after heating the raw starch with a measured amount ofwater, starch granules swell to a pasty substance, regularly arrangedmicelle of starch are broken, crystallites disappear, and the resultingcomposition is easily degraded under the process of amylase.Pre-gelatinized starch is able to swell and/or dissolve in cold or roomtemperature water and form an adhesive paste whose retrogradation islower than that of raw starch, affording easier handling during theproduction process. Raw starch can be pre-gelatinized through solely aphysical modification process without adding any chemical agents andbecomes a hemostatic material with enhanced hydrophilic and adhesiveproperties.

The pre-gelatinized starch of the invention is safe, non-toxic, and hasno adverse side effects. The pre-gelatinized starch is readily degradedand metabolized by enzymes in the body. The described pre-gelatinizedmaterial is safe and biocompatible.

The chemical modifying as described above includes acidolysis,oxidation, esterification, etherification, cross-linking, chemical agentgrafting. Alternatively, multiple modifying processes comprise at leasttwo of the above processes, or one of the above modifying processesperformed at least twice.

According to the present invention, by adding the functional group onthe raw starch glucose units with chemical agents, e.g. by carboxylationmodification, or hydroxylation modification, the starch captureshydrophilic groups in its molecular structure and obtains hydrophilicproperties. By using bifunctional or polyfunctional chemical agents tocross-link the raw starch macromolecules or grafting externalmacromolecurlar hydrophilic groups to the raw starch, the starchacquires enhanced hydrophilic properties and viscosity/adhesiveness in awater solution. The viscosity of modified starch relates to the rawstarch origin and the degree of substitution of external and thecross-linked or grafted functional groups, etc. When contacting blood,the hydrophic and adhesive properties of the prescribed modified starchwill produce a “starch-blood coagulation matrix” with strong adhesivecharacteristics which can seal wounded tissue and stop bleeding. Inaddition, the interaction between the formed blood coagulation matrixand the functional groups of tissue proteins will cause the“starch-blood coagulation matrix” to adhere to and seal the woundedtissue, resulting in hemostasis.

An advantage of the present invention is that the modified starchcompositions are easily swollen and/or dissolved in water, allowing theformed adhesive gel at the wound site to be irrigated and dissolved bynormal saline rinse after hemostasis. Since the residual modified starchparticles not in contact with blood can be easily washed away withwater, absorbed by an aspirator, or wiped away with gauze materials, itwill minimize the residual particles in the wound and the risk of tissueinflammation.

Specifically, the modified starch of the present invention contains oneor more groups of pre-gelatinized starch, acid modified starch, dextrin,oxidized starch, esterified starch, etherified starch, or cross-linkedstarch.

The hemostatic composition of the present invention comprises two ormore modified starches to satisfy the physical and chemical propertiesof a hemostatic agent, where the weight ratio of the two modified starchgroups can be 99:1˜1:99.

Specifically, the weight ratio of the two modified starch groups can be:95:5, 90:10, 85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, or 50:50.

The main physical parameters of the modified starch, according to thepresent invention, are provided below:

The modified starch of the present invention has a molecular weight over15,000 Daltons (for instance, 15,000˜5,000,000 Daltons).

The modified starch of the present invention has a water absorbencycapacity not lower than one time its weight (i.e. 1 gram of describedmodified starch can absorb 1 gram or more of water); whereas it can be1˜500 times generally and 2˜100 times preferably.

The modified starch composition of the present invention comprises, butis not limited to, at least one carboxymethyl starch, hydroxyethylstarch, and cationic starch.

For example, the carboxymethyl starch is a polymer of linear structureas expressed in the following formula:

The modified starches such as carboxymethyl starch (CMS) andhydroxyethyl starch are known clinically as plasma substitutes. Thesemodified starches exhibit biocompatibility and safety with no toxic sideeffects when employed in the human circulatory system. The hemostaticcomposition, according to the present invention, can also include otherplasma substitutes by means of well-known pharmacokinetics approachesand specified physical/chemical properties to produce safe and reliablehemostatic agents.

When cationic starch of the modified starches is used as a hemostaticmaterial, the surface positive charge of the cationic starch can attractand interact with electronegative blood erythrocytes, accelerating theblood coagulation process. Furthermore, when contacting blood, thepositively charged modified starch adheres tightly to tissue, seals thewound, and rapidly stops bleeding. The cationic starch can be usedindependently as a hemostatic material, or mixed with other modifiedstarches as a composite hemostatic material.

The composite modified starch comprises, but not limited to, at leastpre-gelatinized hydroxypropyl distarch phosphate. Specifically, thehydroxypropyl distarch phosphate is produced by cross-linking andetherifying the starch with propylene oxide and phosphoric acid,followed by pre-gelatinization modification through a spray dryingprocess. The hydroxypropyl distarch phosphate has high adhesiveness,strong water absorbency and robust hemostatic effects. It is stable inacidic or alkali environments and can be used as a biocompatible,hemostatic material, a surgical sealant, a tissue healing composition,an anti-adhesion agent, and a tissue repair material.

The cross-linked starch of the present invention comprises, but is notlimited to, at least one epichlorohydrin cross-linked starch and onecross-linked carboxymethyl starch.

The grafted starches of the present invention comprise at least apropylene ester-carboxymethyl starch grafted copolymer and a crylicacid-carboxymethyl starch grafted copolymer. Grafted starch has bothenhanced water absorption capability and high viscosity/adhesiveness.Accordingly, it has a profound effect on hemostasis when applied towound surfaces, especially combat wounds, traumatic wounds, and profusebleeding from large arteries and large veins due to aneurysms or largephlebangioma ruptures.

The modified starch, according to the present invention, can be madeinto powder form, spherical form or aerosol form to be delivereddirectly to the bleeding wound surface.

For large wound surfaces from burns, selecting an aerosol sprayedmodified starch hemostatic powder in combination with a modified starchsponge or film can achieve not only hemostasis but also reduce tissuefluid exudation. This combined application preserves a moist woundsurface and develops a “scaffold” for fiber cell growth and propagationthrough the healing tissue.

Specifically, the hemostatic powder of the present invention is made byan agglomeration process and pellet fabrication. Normally, modifiedstarch grain dimensions are relatively small and light and may need toagglomerate into larger sizes and heavier weights which can readilydisperse into the excess blood and generate coagulation close to thebroken vessels to achieve an optimal hemostatic outcome. Theagglomeration process may not be necessary for large sized modifiedstarch particles such as grafted starch or cross-linked starch.

The modified starch particles of the present invention have a diameterrange of 10˜1000 μm, preferably 30˜500 μm. Starch particles withdiameters of 30˜500 μm represent no less than 95% of the total starchparticles in the preferred embodiment. The measured, optical diameter ofthe starch particles is between 50˜250 μm.

Specifically, because pre-agglomerated modified starch particles aresmall and lightweight, they readily form a colloid on the particlesurface with the moisture in blood. In this case, it affects thehemostatic outcome by preventing water molecules from further dispersingto other starch particles. The present invention accepts and adoptsagglomeration processing technologies in the food and pharmaceuticalindustries to accumulate microscopic modified starch particles in thegeneral 5˜50 μm diameter range, creating clinically applicable particleswith a diameter range of 30˜500 μm. Modified starch particles producedby the process disclosed above exhibit rapid water absorption, stronghydrophilic properties and rapid dispersion in blood to achieve improvedhemostatic outcomes, while not readily forming a colloidal protectinglayer which may disrupt the hemostatic effect.

To fulfill the requirements of clinical operations, the presentinvention provides various methods and processes to produce hemostaticcompositions with acceptable properties that assist doctors withhemostatic therapy during surgery. The powder form modified starchhemostat readily adapts to diffuse oozing of blood on large surfaceareas, and the hemostatic powder can be delivered to a bleeding woundsurface under celioscope, nasoscope, laparoscope or endoscope. Thepowder will have a sealing effect on postoperative biliary fistulas,thoracic cavity fistulas, lymph fistulas, intestinal fistulas, and woundtissue exudation. Excess, residual powder can be rinsed away with normalsaline to reduce the risk of inflammation and infection.

The hemostatic material of the present invention can be a hemostaticsponge, a hemostatic foam, a hemostatic film, or a hemostatic bandage,which can be applied directly to a wound surface to stop bleeding,wherein the hemostatic sponge/foam, the hemostatic film, and thehemostatic bandage can be made into a pad or patch by attaching abacking layer or substrate of fiber fabric.

The hemostatic sponge, hemostatic foam, hemostatic film, and hemostaticbandage according to the present invention can be produced into, but notlimited to, the following forms: columnar, sheet, flocculent, ormembranous.

Concerning active bleeding and high pressure arterial bleeding, surgeonsor emergency responders must apply pressure to the wound to stopbleeding. In this circumstance, the hemostatic powder and the clotformed can be easily washed away by the high pressure blood flowresulting in a failure of hemostasis. In addition, by compressing thehemostatic powder on the bleeding wound, the clotting action forms anadhesive gel which easily sticks and adheres to surgical gloves,instruments, and gauze. As a result, upon removal of gloves, instrumentsor gauze from the coagulated wound, re-bleeding may occur. The presentinvention provides a modified starch sponge or foam which can be appliedand remain directly on the wound, thereby solving the above problem ofre-bleeding. The modified starch hemostatic sponge or foam isabsorbable, easy to use and may remain directly on the wound for asatisfactory effect.

The hemostatic sponge and foam of the present invention can be made fromone or more, but not limited to, modified starch processes such asvacuum freeze drying.

The hemostatic sponge and foam of the present invention can be acomposite hemostatic sponge and composite hemostatic foam made from oneor more compositions of modified starch and other biocompatiblehemostatic materials through, but not limited to, the vacuum freezedrying process.

Whereas, the biocompatible hemostatic materials described abovecomprise, but not limited to, one or more groups of gelatin, thrombin,collagen, carboxymethyl cellulose, oxidized cellulose, oxidizedregenerated cellulose, chitosan, or sodium alginate.

To solve the challenge of molding modified starch into sponges and foam,the present invention combines other known bio-absorbable hemostaticmaterials with strong biocompatibility and clinically acceptablequalities with the modified starch to produce composite hemostaticsponges and foams. Whereas other known bioabsorbable hemostaticmaterials can be of one or more components, modified starches can alsocomprise one or more components, such as modified starch+gelatin,modified starch+collagen, modified starch+thrombin, modifiedstarch+chitosan, modified starch+carboxymethyl cellulose, and modifiedstarch+hyaluronic acid. These combinations can be molded into sponge andfoam form to satisfy clinical requirements.

Weight proportion between the biocompatible modified starch and otherbiocompatible hemostatic materials can be 99.9:0.1˜1:99.

Specifically, the weight ratio between the modified starch and otherbiocompatible hemostatic materials can preferably be: 95:5, 90:10,85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, 50:50, 45:55, 40:60,35:65, 30:70, 25:75, or 20:80.

This additional coagulant material can be added to the modified starchhemostatic sponge or foam of the present invention directly during thevacuum freeze drying production process to produce a compositehemostatic sponge or composite hemostatic foam. The production processcan, but not limited to, pre-mixing the coagulant material with themodified starch directly before vacuum freeze drying process.

Whereas the coagulant of the present invention comprises, but notlimited to, one or more combinations of the following blood coagulationfactors: thrombin, fibrin, calcium agent, polypeptide, peptide, aminoacid, and protamine.

The modified starch sponge and foam of the present invention can bemanufactured into hemostatic sponge and foam form by a vacuum freezedrying process utilizing a forming agent or a plasticizing agent.

Whereas the forming agents as described above comprise, but not limitedto, organic forming agents, inorganic forming agents, natural formingagents, and man-made plasticizing agents, which can include, but notlimited to, one or more combinations of glycerol, kaolin, sorbitol,ethanol, ammonia, and polyethylene glycol.

Specifically, vacuum freeze drying process is a drying method thatfreezes wet material or solutions to a solid state under lowtemperatures (−10˜−50° C.) and then converts the solid material into agas and then, in a vacuum (1.3-1.5 Pa), back to a solid material withoutan intermediate liquid phase (sublimination). As the vacuum freezedrying is processed under low temperature and low pressure, the moisturesublimes directly to produce a substance with numerous specialproperties.

The basic parameters of the vacuum freeze drying process specify bothphysical parameters and process parameters. The physical parametersinclude thermal conductivity, transfer coefficient, etc. The processparameters include freezing, heating, state of the material, etc.Continued research on this freezing process involves experiments toidentify the optimal freezing curve.

As the described biocompatible, modified starch hemostatic material canbe made into a hemostatic glue, the physical form can be colloidal,dissolved colloidal, thawed colloidal, semi-liquid or gelatinous, etc.

The hemostatic glue can be produced by adding other liquids, not limitedto water, to the modified starch by diluting, swelling, or dissolvingthe liquids in certain proportions.

The present invention discloses the topical application of modifiedstarch as a hemostatic agent to manage and control bleeding woundsurfaces in humans, mammals, birds, or reptiles, and the internalapplication for hemostasis of bleeding wound surfaces within humanbodies, tissues and organs following surgical operations and traumatreatments, under open surgery or nasoscope, laryngoscope, endoscope andlaparoscope.

It is important to note that the modified starch hemostatic composition,according to the present invention, can be applied for hemostasis onbleeding bone tissue caused by surgery or trauma, particularly forhemostasis in spongy bone tissue. In thoracic or neurological operationsinvolving some patients, such as children, elderly people, and patientswith osteoporosis, sternal bleeding and skull bleeding is difficult tocontrol. It is common to apply bonewax to the sternum or skull, however,bonewax is slow to absorb and may cause complications such as non-unionor infection. The modified starch composition, according to the presentinvention, is a biocompatible substitute for bonewax to control bonebleeding and mechanically seal the wound caused by surgery or traumawith robust hydrophilic properties, strong adhesiveness and ease ofmolding. The modified starch hemostatic material degrades rapidly aftersurgery and avoids the complicating issues of non-union and infectionassociated with bonewax.

As the modified starch is applied as hemostatic material, otherbiological benefits of the modified starch are worthy of attention. Itis essential to evaluate the modified starch composition for itspositive, additional effects on wound inflammation, tissue adhesion andtissue healing while it functions as a hemostat.

Through research and experimentation, it is proven this modified starchhemostatic material, according to the present invention, has furtherapplication as an absorbable, postoperative tissue adhesion barrier. Theadhesion barrier of the modified starch, according to the presentinvention, prevents wounded tissue or organs from adhering to othertissue or organs in the vicinity, thereby reducing local bleeding andexudation, by mechanically isolating the wound or wound surface fromadjacent tissue and surrounding organs, such as the peritoneum.

The modified starch, according to the present invention, can promotetissue healing, including skin, subcutaneous soft tissue, musculature,bone tissue, neurological tissue, nerve tissue, and wounded tissue ofthe liver, kidney, spleen, etc. through proper dosage and application.The modified starch can create a “scaffold” for skin tissue cell healingand growth of skin tissue from large wound surfaces due to burns, and a“scaffold” for osteocyte growth and propagation in bone defects fromtrauma, bone tumor resection, etc.; and a “scaffold” for neurologicaltissue cell growth and propagation when applied to injured neurologicaltissue caused by brain trauma, brain tumor resection, etc.

The mechanism for promoting tissue healing is the “glue” formation aftermodified starch contacts blood and establishes the “scaffold” on thewound surface which facilitates the adherence, growth, connection andpropagation of tissue cells such as osteoblasts or fibroblasts. Inaddition, local blood platelets are increasingly concentrated on thewound and, when activated, release tissue factors which promote healing.

The modified starch, according to the present invention, has furtherapplications as a biocompatible surgical sealant, capable of forming aprotective layer of colloid or film on the wound surface to seal andprevent drainage of blood, tissue fluids, lymph fluid, cerebrospinalfluid, bile, gastric fluid, or and other intestinal fluids resultingfrom surgery and trauma treatment. This sealing effect will preventlymph fistula leakage, bile flaccidity, pleural flaccidity, intestinalflaccidity, cerebrospinal flaccidity, and vascular flaccidity.

The modified starch, according to the present invention, has furtherapplications as a biocompatible tissue adhesive, capable of adhering,repairing, and bonding wounded nerve tissue, musculature, and tissues ofthe bone, skin, viscera, and subcutaneous tissue. It will also bondother curative materials to wounded tissue and organs.

The differences between the present invention and prior hemostaticmaterials are:

In contrast, the microporous polysaccharide hemospheres of U.S. Pat. No.6,060,461, an absorbable, biocompatible hemostatic material, are formedby cross-linking starch with epichlorohydrin. The mechanism of themicroporous polysaccharide hemospheres involves the molecular sieving ofblood components based on their molecular weight. This microporoushemostat allows water molecules and other lower weight molecules intoits particles and concentrates heavier molecules, such as erythrocytes,platelets, and fibrinogen, on the surface of the particles to promoteblood coagulation.

The microporous polysaccharide hemospheres of U.S. Pat. No. 6,060,461are produced under a proprietary process not disclosed in the patent.However, normal modified starches, including cross-linked modifiedstarch, do not necessarily possess the microporous structure of themicroporous polysaccharide hemospheres as described in U.S. Pat. No.6,060,461. By contrast, the present invention does not employ themicroporous properties of a molecular sieve in modified starch toachieve hemostasis. Rather, the present invention adopts other technicalprocesses to import hydrophilic groups to raw starch molecules whichenable the modified starch to directly interact with water moleculesunder hydration, resulting in rapid dehydration of blood andconcentration of blood clotting components. This action does not relateto whether or not the modified starch has a microporous surface.

In addition, by means of changing the degree of substitution, selectingproportional ratios of amylopectin and amylose, adding functional groupsto starch molecular or modifying the functional groups in starch chains,etc, the present invention is able to increase the hydrophilic propertyand viscosity of the modified starch, which produces an “adhesive gel”that adheres strongly to tissue and mechanically seals broken bloodvessels after contact with blood. The above properties were notidentified in the microporous polysaccharide hemospheres of U.S. Pat.No. 6,060,461 and these described properties in present invention haveadvantages over prior hemostatic materials. The modified starch in thisinvention can be made into hemostatic powder, hemostatic sponges andfoam, hemostatic glue and gel, independent of whether or not themodified starch has a microporous structure on the surface. On thecontrary, the hemostatic effect of the powder, the sponge, the foam, theglue and the gel relates to the induced physical and chemical propertiesof the modified starch under which the invention is synthesized.

When compared with Chinese publication patent number CN1533751A,hemostatic dressings consist of two necessary components: fabric and amulti-porous polymer substrate attached to the fabric, which then formsa composite structure. The fabric is made from the oxidized regeneratedcellulose described previously. Since human physiology lacks sufficientdegrading enzymes for oxidized regenerated cellulose, this hemostaticmaterial is slow to absorb in the human body, thus risking localinfections and compromising tissue healing In the description of thesubstrate, the patent identifies dextran and carboxymethyl cellulose asderivatives of dextran. Cellulose and starch are two large classes ofdextran with distinctly different properties, though both arepolysaccharide dehydrating and poly-condensing from glucose monomers.Firstly, the polymerization degree of starch is generally from hundredsto thousands, and the molecular weight of starch is from tens ofthousands of Daltons to hundreds of thousands of Daltons. Thepolymerization degree of cellulose is generally thousands and themolecular weight of cellulose is from hundreds of thousands of Daltonsto millions of Daltons. Secondly, all repeating glucose chains in starchare arranged in the same direction, while repeating cellulose chainsconnect to each other by rotating 180° along the axial direction, whichproduces the different glucose and cellulose structures. Further, starchis easily degraded and metabolized by amylase and carbohydrase, enzymesabundant in the human body. Conversely, cellulose is slow to metabolizeand absorb since the human body lacks sufficient amounts of therequisite degrading enzymes.

Consequently, when producing biocompatible, absorbable materialsemployed for hemostasis, the properties of starch are superior to thoseof cellulose since modified starch is readily degraded into glucose bythe abundant amount of amylase in the human body and absorbed rapidly.The biocompatibility advantage of starch over cellulose is apparent.Furthermore, the oxidized regenerated cellulose dressing has weakadhesion to tissue, requires compression to maintain contact withtissue, and therefore cannot effectively seal blood vessels in thewounded surface, limiting its clinical applications.

In addition to the above different properties, the modified starchdescribed in the present invention has a bacterio-static andanti-inflammatory effect on bleeding wound surfaces. The modified starchhemostatic material, according to the present invention, has ahemostatic effect which controls bleeding, reduces blood and tissuefluid exudation, and maintains a moist wound surface. As a result, itsuppresses the growth of bacteria and reduces the inflammatory response,diminishing local irritation and relieving pain. Furthermore, tostrengthen the anti-inflammatory response, known antibiotic or otheranti-pyrotic agents can be added to the modified starch during themanufacturing process to produce hemostatic powder, sponges and foams,hemostatic glues and gels, etc., all of which are suitable for topicaland internal clinical applications.

Another advantage of the modified starch hemostatic material accordingto the present invention is the rapid particle dissolution in water,facilitating the easy removal of excess modified starch particles fromthe wound by simple saline irrigation. The residual modified starch notactively involved in hemostasis can be rinsed away by irrigation. In thetreatment of battle wounds, self rescue, or first aid, the hemostaticmaterial remaining in small amounts will be absorbed by the body and theirritation wound debridement or gauze removal is avoided.

The modified hemostatic starch material has properties of stability,extended shelf life, resistance to high and low pressure, resistance tohigh (up to 60° C.) and low (down to −40° C.) temperatures, storageconvenience, and physical stability. Therefore, it can be employed as ahemostatic material for military, emergency, and first-aid uses.Particularly, it can be adapted in extreme environmental conditions suchas desert areas, polar regions, alpine areas, outer space, andunderwater probes. The modified starch sponge has the physicalproperties of pliability, flexibility, moldability, and curling. It canbe adapted for wound surfaces with various shapes, sizes, and features,such as deep and irregular anatomical wounds and organs, both inside andoutside the lacuna surfaces, and may be applied via endoscope,laparoscope or open surgery.

To enhance the safety of applying the modified starch to wound surfaces,tissue, etc., the modified starch material, according to the presentinvention, can be packaged and sterilized with, but not limited to,gamma irradiation, oxirane, and ozone sterilization.

However, adopting alcohol sterilization, autoclave or steamsterilization is not recommended as it may change the physical andchemical properties of the modified starch and compromise its hemostaticeffect.

A. Preparation of Modified Starch Powder Preferred Embodiment 1

A biocompatible modified starch material uses as hemostatic material. Itincludes carboxymethyl starch (66#). Carboxymethyl starch material isadded into an agglomerator at 40˜50° C. Distilled water is added. Theprocesses include particles coagulation (agglomeration) and pelletmaking. Molecule weight of the carboxymethyl starch (66#) is15,000˜2,000,000 Dalton. Diameter of the particles is 10˜1000 μm.Particle diameter is between 30 and 500 μm in no less than 95% of theparticles. Viscosity of a 6.67% suspension at 37° C. is 557.9 mPa·s.Work of adhesion under room temperature is 68.1 g·sec when the modifiedstarch is saturated with water.

Preferred Embodiment 2

A modified starch absorbable hemostatic material includes hydroxyethylstarch (88#). Hydroxyethyl starch material and distilled water are addedinto an agglomerator 40˜50° C. The processes include particlescoagulation (agglomeration) and pellet making Molecule weight of thehydroxyethyl starch (88#) is 15,000˜1,000,000 Dalton. Diameter ofhydroxyethyl starch (88#) particles is 10˜1000 μm. Particle diameter isbetween 50 and 500 μm in no less than 95% of the particles. Work ofadhesion under room temperature is 420.9 g·sec when the modified starchis saturated with water.

The water absorption ability of this invention is measured by acapillary method. Water is added into an acid burette so that the liquidlevel at zero graduation of the acid burette equals to the bottom of thefilter plate of a sand core funnel. Filter paper is trimmed into a discwith a 2.25 cm radius, weighed, and put into the sand core funnel untilit fully touches the filter plate. The piston is opened until the filterpaper is fully absorbed with water. The acid burette is adjusted to zerograduation. 0.1 g sample powder is weighed, scattered evenly on thefilter paper, and placed in the sand core funnel. Starting from the timewhen liquid begins to fall, liquid level is recorded every 20 s, 40 sand 60 s. Water absorption speed and water absorption saturation perunit time are calculated.

A comparison of water absorption ability between carboxymethyl starch66# and Arista™ groups is illustrated in Table 1.

As illustrated in Table 1, the water absorption speed of carboxymethylstarch 66# within all three 20 s intervals are greater than that ofArista™, indicating that 66# absorbs water faster than Arista™ and ismore effective. The capacity of water absorption of 66# is almost 5times as that of Arista™ in the first 20 s interval.

The water absorption speed refers to the average water absorption speedin the first 20 s, the second 20 s, and the third 30 s intervals. V20s=water absorbed in 20 s(ml)/20 (s).

Saturation rate of water absorption refers to the amount of waterabsorbed by the sample in a certain period of time divided by itsmaximal water absorption capacity (i.e. the absolute value of waterabsorbency). This measure reflects the water absorption speed of thesame sample from another angle.

As illustrated in Table 1, 66# has a higher saturation rate of waterabsorption than Arista™ at the 20 s, 40 s, and 60 s intervals,indicating that 66# absorbs more water than Arista™ in a same period oftime. For 66#, 58 percent of total water absorbency is achieved in 20 s;nearly 95% is achieved in one minute. The water absorption speed of 66#is greater than that of Arista™.

The stickiness of the present invention is measured as the work ofadhesion using a texture analyzer (physical property analyzer; StableMicro System, Model TA-XT plus). Probes used in the experiment includesA/BE (backward extrusion probe) and P36R (cylindrical probe).

Experiment conditions are: pre-experimental speed: 0.5 mm/sec;experimental speed: 10.0 mm/sec; stress: 100 g; retrieval distance: 5.0mm; contact time: 10.0 sec; trigger: automatic, 5 g.

A comparison of work of adhesion among carboxymethyl starch 66#(according to the present invention), hydroxyethyl starch 88#, andArista™ is illustrated in Table 2.

When the probe moves back, it will encounter the adhesive force producedby the sample. For the probe to separate completely from the sample, itmust do work. The work done during this period is referred to as thework of adhesion and can be used to measure the adhesive strength(degree of firmness) between the adhesive agent and probe surface.

25% saturation rate refers to the saturation at ¼ maximal waterabsorption capacity.

50% saturation rate refers to the saturation at ½ maximal waterabsorption capacity.

100% saturation rate refers to the saturation at maximal waterabsorption ability.

As illustrated in table 2, the adhesiveness (stickiness) of Arista™ ismuch lower than that of 66# and 88#. The work of adhesion of 88#decreases with increasing saturation rate, and has particularly highadhesiveness (stickiness) and at lower saturation rate. Adhesiveness(stickiness) of 66# increases gradually. At maximal saturation, bothmaterials have significantly higher stickiness than Arista™, and producebetter effects of adhesive plugging.

Viscosity of the present invention is measured by a viscometer(brookfiled Dv-2). Test conditions are: Rotor 3; rotating speed: 60;concentration of modified starch solution: 6.67%; temperature: 37° C.

A comparison of viscosity between carboxymethyl starch 66# in thepresent invention and Arista™ is illustrated in Table 3.

As illustrated in Table 3, the viscosity of 66# is significantly higherthan that of Arista™.

Preferred Embodiment 3

A biocompatible modified starch material uses as hemostatic material. Itincludes pre-gelatinized hydroxypropyl distarch phosphate (51#). Itsmolecule weight is over 15,000 Dalton (15,000˜2,000,000 Dalton).Diameter of its particles is 10˜1000 μm. Particle diameter is between 50and 500 μm in no less than 95% of the particles.

Preferred Embodiment 4

A biocompatible modified starch applied in hemostasis includescrosslinked carboxymethyl starch (66#+). Its molecule weight is over15,000 Dalton (15,000˜2,000,000 Dalton) and the diameter of particles is10˜1000 μm; among them, those with diameters between 50 and 500 μm takeno less than 95% of total amount of starch particles.

Preferred Embodiment 5

A biocompatible modified starch uses as hemostatic material. It includespre-gelatinized starch prepared using a spray drying process. Itsmolecule weight is over 15,000 Dalton (15,000˜2,000,000 Dalton).Diameter of particles is 10˜1000 μm, Particle diameter is between 50 and500 μm in no less than 95% of the particles.

The water absorbency of various modified starch is determined bycentrifugation and the results are illustrated in Table 4:

Water absorbency refers to the maximal water that 1 g sample can absorb.Water absorbency (ml/g)=amount of absorbed water (ml)/amount of sample(g).

As illustrated in table 4, all prepared modified starches have betterwater absorbency.

Control Experiment 1

Hemostatic effects in a liver bleeding model in New Zealand rabbits.

Objective: To investigate the hemostatic effects of 66# products in aliver bleeding model in New Zealand rabbits.

Test Drugs:

Name: Product 66# (carboxymethyl starch hemostatic spheres)

Animals: New Zealand white rabbits, supplied by Laboratory AnimalCenter, the Second Military Medical University.

Certification Number: SCKK (SH) 2002-0006

A total of 15 animals were used (n=5 per group). Body weight: 2.0±0.3kg.

Methods:

New Zealand white rabbits were randomly divided into 3 groups, a product66# group, a positive control group (Arista™) and a negative controlgroup (raw starch) (n=5, respectively). The rabbits were anaesthetizedwith sodium pentobarbital (40 mg/kg) via ear vein injection. Rabbitswere fixed in a supine position. Hair was removed. After disinfection,the abdominal cavity was opened layer by layer to expose the liversufficiently. A 1 cm diameter and 0.3 cm deep wound was produced using apuncher on the liver surface. Hemostatic material was appliedimmediately. The wound was pressed for 20 s. Hemostatic effects wereobserved in each animal group. The animals received Arista™ and rawstarch in the positive control group and the negative control group,respectively. All the animals received un-restricted food and waterafter the surgery. At half an hour, one, two, three and seven days aftersurgery, one animal from each group was chosen and received anesthesia.The hepatic wound surface was stained with iodine to observe thedegradation of the hemostatic materials. The wound tissue was taken outand fixed with 10% formaldehyde. Tissue sections were prepared and usedto observe the degradation of the hemostatic materials.

Dosage: 50 mg/wound surface

Route: spray applying

Frequency: once/wound surface

Outcomes and observation time: hemostatic effect, absorption anddegradation, healing of the wound surface. Observation times were: halfan hour, one day, two days, three days and seven days after the surgery.

Results:

Effects on Hemostasis

In the positive control group (Arista™), the bleeding was stoppedimmediately after the hemostatic material was sprayed on. In the product66# group, the bleeding was also stopped immediately. In the raw starchgroup, the bleeding could not be stopped after the hemostatic materialwas sprayed on even with wound pressing. (See FIGS. 1 to 3)

Degradation In Vivo

There was no iodine color reaction in the positive control group(Arista™) and product 66# group at half an hour. Color reaction waspositive in the negative control group at half an hour later, but not 24hours later.

Control Experiment 2

Degradation in Abdominal Cavity of Mice

Objective: To investigate the adhesion and degradation of product 66# inabdominal cavity of mice.

Test Drugs:

Name: Product 66# (carboxymethyl starch hemostatic spheres)

Animal: ICR mouse, supplied by Laboratory Animal Center, the SecondMilitary Medical University.

Certification Number of Animals: SCXK (SH) 2002-0006

A total of 30 animals were used (n=10 per group). Body weight: 18-23 g.Half were females and half were males.

Methods: Product 66#, the positive control Arista™, and the negativecontrol raw starch, were prepared into 0.1 g/ml solutions with normalsaline. A total of 30 ICR mice were randomly divided into three groups,a product 66# group, a positive control group (Arista™) and a negativecontrol group (raw starch). Animals received intraperitoneal injectionof 1 ml of the above-mentioned solutions. Twenty-four hours later,abdominal cavity was opened. Iodine was applied in the abdominal cavityto observe color change and visceral adhesion. Animals in the positivecontrol group and the negative control group received Arista™ and rawstarch, respectively.

Dosage: 1 ml/per mouse

Route: intraperitoneal injection

Frequency: once/per mouse

Outcomes and observation time: The abdominal cavity was opened 24 hoursafter the administration to observe organ adhesion and materialdegradation.

Results:

Adhesion In Vivo

Twenty-four hours later, no organ adhesion was found in the abdominalcavity in 66# experimental group. (See FIG. 4)

Degradation In Vivo

No iodine color reaction was found in 66# experimental group attwenty-four hours later, indicating that 66# had been degradedcompletely within the mice body. (See FIG. 5)

Control Experiment 3

An investigation of hemostastic effects of products 66# and 88# in acanine femoral artery bleeding model.

Objective: To observe the hemostatic effect of product 66# and 88# inserious trauma, and compare the hemostatic effect of product 66# andproduct 88# with Arista™.

Animals: experimental dogs.

A total of 20 male animals were used (n=5 per group). Body weight: 20-25kg.

Methods: The animals were randomly divided into a control group(pressing with gauze), a product 66# group, a product 88# group, and anArista™ group. The femoral artery was exposed and punctured with a No.18 needle (diameter: 2F). Blood was allowed to flow freely for 2 second.After establishment of the canine femoral artery injury model, product66#, 88#, or Arista™ (1 g, respectively) was applied on the bleedingsites and pressed manually. Animals in the control group receivedpressing with gauze. Then, after pressing for 60 s, 90 s, 120 s, and 180s, hemostatic effects of the materials were observed. Successful caseswere recorded. Stop of bleeding or blood oozing at the puncturing wasused as a criterion for successful hemostasis. Hemostatic status ofexperimental animals under different hemostatic conditions is shown inTable 5.

Conclusion

Product 66#, 88# and Arista™ had significant hemostatic effects oncanine femoral artery bleeding as compared with the control group.Product 66# and 88# had better sealing effects on the punctured site atfemoral artery and significantly shorter hemostatic time than Arista™.Furthermore, the more adhesive product 88# had better sealing effects onthe punctured site at femoral artery and shorter hemostatic time thanproduct 66# and Arista™.

Control Experiment 4

An investigation on postoperative intestinal adhesion in rats.

A sample of product 66# in the experimental group was compared with apositive control of sodium hyaluronate on sale for medical use, and ablank control.

Experimental Animals and Grouping

Thirty-four male SD rats weighing 200˜250 g were supplied by LaboratoryAnimal Center, the Fourth Military Medical University. They were dividedinto three groups: a blank control group, a 66# group, and a positivecontrol group of medical sodium hyaluronate (SH). Each group consists of11 or 12 rats.

Preparation of the Rat Intestinal Adhesion Model

Animals in all groups were fasted but received unrestricted supply ofwater 12 hours before the operation. Rats were anaesthetized with 3%sodium pentobarbital (30 mg/kg; intramuscular). The cecum was exposedthrough a 2 cm midline incision in the lower abdomen. The serosa of thececum was scraped until blood oozed. Anhydrous ethanol was applied onthe wound surface. The mesenteric artery of cecum was clamped with a5-finger clamp for 2 min to induce temporary ischemia. After thesetreatments, the wound surface in the 66# group and SH group were coveredwith the corresponding drugs. The blank controls did not receive anydrug. The cecum was placed back into the original position in theabdominal cavity. The opposing abdominal wall was damaged with ahemostatic forcep and the abdominal cavity was closed with No. 1-0thread layer by layer. Rats received intramuscular gentamicin injection(4U) for 3 consecutive days after the surgery to prevent infection.Fourteen days later, the same method of anesthesia was used to open theabdominal cavity for examination and sampling.

Relevant Measurements

1) General Condition: Survival of the rats was recorded after theoperation.

2) Intestinal Adhesion: The abdominal cavity was opened using a U-shapedincision (bottoms down) that included the original midline incision. Thetissue flap was lifted up to expose the abdominal cavity. Adhesionbetween the cecum and the abdominal wall was observed and graded usingthe Nair 5 Grading System: Grade 0: no adhesion; Grade 1: one adhesionbelt between the viscera or between different points of the abdominalwall; Grade 2: two adhesion belts between the viscera or between theviscera and abdominal wall; Grade 3: two adhesion belts; no directadhesion of the viscera to the abdominal wall; Grade 4: the visceraadhere to the abdominal wall directly, regardless of the number ofadhesion belt.

Please refer to FIG. 6 for the intestinal adhesion in the blank controlgroup. FIG. 7 illustrates the effects of 66# in preventing intestinaladhesion. FIG. 8 illustrates the effects of sodium hyaluronate inpreventing intestinal adhesion. These results indicated that the sodiumhyaluronate and carboxymethyl starch 66# could significantly reducepostoperative intestinal adhesion in rats.

Control Experiment 5

An investigation on the postoperative bone healing condition in rabbits

Experimental Method

Major Materials

Carboxymethyl starch 66#, pre-gelatinized hydroxypropyl distarchphosphate 51#, bone wax, and blank control group.

Experimental Animals and Grouping

32 New Zealand adult female rabbits, 2.0˜2.5 kg, were supplied byLaboratory Animal Center of the Fourth Military Medical University. Twodefect pores could be drilled on each animal. The rabbits were randomlydivided into a blank control group, a 66# group, a 51# group, and a bonewax group (n=8, respectively).

Operative Method

Animals were anaesthetized with 3% sodium pentobarbital via the ear veininjection (30 mg/kg) and fixed on a prone position on an operativetable. A 4 cm sagital incision was made along the midline to expose theskull. The periosteum was removed completely. A round defect pores wasmade on each side of cranial midline with a 6 mm diameter drill bit(diameter: 6 mm) The defects spanned the layer of the skull (Thethickness of the skull is essentially uniform in parietal bone). Themidline was not crossed. The defects were covered randomly with one ofthe aforementioned materials. No material was applied in the controlgroup. The periosteum and scalp were sutured with absorbable 4-0 thread.The wound was aseptically dressed. Animals were placed back to the homecage and raised for 6 weeks. Animals received intramuscular gentamicininjection (4U) 3 consecutive days after the surgery to preventinfection. The general situation of the animals was monitored everyday.

Seven days prior to the sacrifice, animals received calcein (Sigma,dissolved in 2% sodium bicarbonate) via the ear veins (20 mg/kg). At 1day to the sacrifice, animals received tetracycline (30 mg/kg; Sigma,dissolved in double distilled water) via the ear vein on the other side.Calcein and tetracycline were deposited on the mineralizing front ofnewly formed bone matrix, thus could be used as markers to measure thegrowth range of the bone during the six days.

Sampling and the Assessment for Bone Healing

1. Sampling: Six weeks after the operation, the animals were sacrificedwith excessive intravenous pentobarbital injection. The skull coveringat least 1.5 cm from the defect edge was included. The samples includedperiosteum and cerebral dura mater. The samples were fixed with 70%alcohol.

2. Bone Healing Score: Bone healing of all defects was assessed withhealing score. The standards were: 0=no visible defect; 1=a few visibledefects; 2=moderate visible defects; 3=extensive visible defects.

3. Pathology and Immunohistochemistry: Fixed skull sample was embeddedwith paraffin. Sections were prepared using routine methods, observedand photographed using an ultraviolet fluorescent microscope. Thefluorescent markers, calcein and tetracycline, bind to the newly formedbone matrix and that not calcified yet, respectively, thus showinglinear fluorescence. The distance between the two fluorescent labelinglines indicates mineral apposition rate (MAR) during the 6 days andactivity of osteoblasts, (osteogenetic speed).

${MAR} = \frac{{Distance}\mspace{14mu}{between}\mspace{14mu}{two}\mspace{14mu}{fluorescent}\mspace{14mu}{labeling}\mspace{14mu}{lines}\mspace{14mu}({\mu m})}{{Days}\mspace{14mu}{between}\mspace{14mu}{two}\mspace{14mu}{administrations}}$

Sections were deparaffinated, dehydrated, transparentized and stainedwith Goldner-Mason-Trichrome and ponceau. The osteoid area andmineralization bone area were labeled in different colors, and wereobserved under a light microscope, photographed. Areas stained withdifferent materials were analyzed with image analysis software.

$\mspace{20mu}{{{Osteoid}\mspace{14mu}{Rate}} = \frac{{Osteoid}\mspace{14mu}{area}\mspace{14mu}{of}\mspace{14mu}{defect}\mspace{14mu}{pore}}{{Area}\mspace{14mu}{of}\mspace{14mu}{defect}\mspace{14mu}{pore}}}$${{{Min}{eralization}}\mspace{14mu}{Bone}\mspace{14mu}{Rate}} = \frac{{Mineralization}\mspace{14mu}{bone}\mspace{14mu}{area}\mspace{14mu}{of}\mspace{14mu}{defect}\mspace{14mu}{pore}}{{Area}\mspace{14mu}{of}\mspace{14mu}{defect}\mspace{14mu}{pore}}$$\mspace{20mu}{{{Defect}\mspace{14mu}{Area}\mspace{14mu}{Rate}} = \frac{{congenital}\mspace{14mu}{absence}\mspace{14mu}{area}\mspace{14mu}{of}\mspace{14mu}{defect}\mspace{14mu}{pore}}{{Area}\mspace{14mu}{of}\mspace{14mu}{defect}\mspace{14mu}{pore}}}$

4. Evaluation Criteria: Bone healing score, mineral deposition rate, theosteoid area, mineralization bone area, and congenital absence area.

5. Statistical Analysis

Data were treated with SPSS 11.0 statistical software. Analysis ofvariance was employed in the comparison of data among the groups.

Experimental Results

Result

The healing score of defects in 51# group and 66# group wassignificantly lower than that in the blank control group and the bonewaxgroup at the sixth week after the operation. The mineral depositionrate, osteoid area, mineralization bone area and other indexes in 51#group and 66# group were significantly higher than those in the blankcontrol group, whereas the congenital absence area in 51# group and 66#group was significantly lower than that in the blank control group.

As shown in FIG. 9, a representative photograph for bone healing indexesin rabbits, 66# and 51# had remarkable effects in improving the skullhealing in rabbits.

B. Modified Starch Sponge Preferred Embodiment 6

Two gram pre-gelatinized hydroxypropyl distarch phosphate 51# is addedinto 30 ml water and stirred continuously to make starch particles swellsufficiently and disperse into a uniform suspension. Several drops ofglycerol are added as a plasticizing agent (forming agent). The liquidis then put in a container and pre-cooled at −40° C. for 22 hours. It isthen frozen and dried for 20 hours at −40° C. in a vacuum <20 Pa in afreezing drier. The final product is modified starch compositehemostatic sponge A.

Preferred Embodiment 7

One gram pre-gelatinized hydroxypropyl distarch phosphate 51# is addedinto 30 ml water and stirred continuously to make starch particles swellsufficiently and disperse into a uniform suspension. The liquid is thenput in a container and precooled at −40° C. for 22 hours. It is thenfrozen and dried for 20 hours at −50° C. in a vacuum <20 Pa in afreezing drier. The final product is modified starch compositehemostatic sponge B.

Referring to FIG. 10, a scanning electron microscope photo for thesection of hemostatic sponge A is illustrated, and referring to FIG. 11,a scanning electron microscope a photo for the section of hemostaticsponge B is illustrated. Adding plasticizing agent during production canreduce sponge pore's diameters and enhance its density and specificsurface area.

Preferred Embodiment 8

Two gram carboxymethyl starch 66# is added into 30 ml water and stirredcontinuously to make starch particles swell sufficiently and disperseinto a uniform suspension. The liquid is then put in a container andprecooled at −40° C. for 22 hours. It is then frozen and dried for 20hours at −50° C. in a vacuum <20 Pa in a freezing drier. The finalproduct is modified starch composite hemostatic sponge C.

Preferred Embodiment 9

Three gram crosslinked carboxymethyl starch 66#+ is added into 30 mlwater and stirred continuously to make starch particles swellsufficiently and disperse into a uniform suspension. The liquid is thenput in a container and precooled at −40° C. for 22 hours. It is thenfrozen and dried for 20 hours at −45° C. in a vacuum <20 Pa in afreezing drier. The final product is modified starch compositehemostatic sponge D.

Preferred Embodiment 10

Three gram hydroxyethyl starch 88# is added into 30 ml water and stirredcontinuously to make starch particles swell sufficiently and disperseinto a uniform suspension. The liquid is then put in a container andprecooled at −40° C. for 22 hours. It is then frozen and dried for 20hours at −50° C. in a vacuum <20 Pa in a freezing drier. The finalproduct is modified starch composite hemostatic sponge E.

Preferred Embodiment 11

A certain amount of medical gelatin (10 g) is added into 100 ml waterand heated in a beaker to 60° C. to form a colloidal solution. Two gramcarboxymethyl starch 66# is added into 30 ml water and stirredcontinuously to make starch particles swell sufficiently and disperseinto a uniform suspension. The two solutions are mixed together in acontainer with the mass ratio of medical gelatin to 66# at 1:1. Afterprecooling at −40° C. for 22 hours, it is frozen and dried for 20 hoursunder −45° C. in a vacuum <20 Pa in a freezing drier. The final productis modified starch composite hemostatic sponge F.

Preferred Embodiment 12

A certain amount of medical gelatin (10 g) is added into 100 ml waterand heated in a beaker to 60° C. to form a colloidal solution. Two gramcarboxymethyl starch 66# is added into 30 ml water and stirredcontinuously to make starch particles swell sufficiently and disperseinto a uniform suspension. The two solutions are mixed together in acontainer with the mass ratio of medical gelatin to 66# at 2:1. Afterprecooling at −40° C. for 22 hours, it is frozen and dried for 20 hoursunder −45° C. in a vacuum <20 Pa in a freezing drier. The final productis modified starch composite hemostatic sponge G.

Preferred Embodiment 13

A certain amount of medical gelatin (10 g) is added into 100 ml waterand heated in a beaker to 60° C. to form a colloidal solution. One gramhydroxypropyl distarch phosphate 51# is added into 30 ml water, andstirred continuously to make starch particles swell sufficiently anddisperse into a uniform suspension. The two solutions are mixed togetherin a container with the mass ratio of medical gelatin to hydroxypropyldistarch phosphate 51# at 2:1. After precooling at −40° C. for 22 hours,it is frozen and dried for 20 hours under −45° C. in a vacuum <20 Pa ina freezing drier. The final product is modified starch compositehemostatic sponge H.

Preferred Embodiment 14

A certain amount of medical gelatin (10 g) is added into 100 ml waterand heated in a beaker to 60° C. to form a colloidal solution. One gramhydroxypropyl distarch phosphate 51# is added into 30 ml water andstirred continuously to make starch particles swell sufficiently anddisperse into a uniform suspension. The two solutions are mixed togetherin a container with the mass ratio of medical gelatin to hydroxypropyldistarch phosphate 51# at 1:1. After precooling at −40° C. for 22 hours,it is frozen and dried for 20 hours under −45° C. in a vacuum <20 Pa ina freezing drier. The final product is modified starch compositehemostatic sponge I.

0.1 g of the above-mentioned sponges is used to compare the chemical andphysical property. Results are shown in Table 8.

An introduction to the measurement of contact angle

Apparatus: OCA40 Micro video contact angle measuring system(Dataphysics, Germany)

Methods: A sessile drop method was used to track and record waterabsorbing status of sponges using dynamic recording function and camerafunction. Details of the procedure were: a sponge sample was placed onan object table, and adjusted slowly to make the object stage appear atthe inferior ⅓ portion of the visual field. A needle filled withdeionized water connected an injection unit was used. A drop of waterwith a certain volume was suspended at the tip of the needle using anautomatic injection system. The system was focused so that the image ofthe sponge sample and water drop appeared clearly in the visual field.The object table was raised slowly so that the sponge sample touched thewater drop. The camera function and dynamic recording function wereturned on simultaneously to observe the process of water drop beingabsorbed and obtain the dynamic contact angle values.

The water absorption ability of composite hemostatic sponges is shown inTable 9.

Water absorbency of the sponges is determined by centrifugation. 0.025 gsponge was placed in 2 ml water, equilibrated for 10 minutes, andcentrifuged for 10 minutes at 500 rpm. Sample was taken out, andweighed. The amount of remaining residual liquid was calculated. Eachsample was measured 6 times. Average values were used.

Volume density of sponges was measured. A sponge sample was cut intocertain length and width and height, and weighed to calculate thedensity.

Hygroscopicity and water absorbency of the sponges were observed throughthe OCA40 Micro video contact angle measuring system of Dataphysics,Germany.

A comparison of water absorbency between composite hemostatic spongesand other hemostatic sponges is shown in Table 10.

As shown in Table 10, composite hemostatic sponge containing modifiedstarch had significantly higher water absorbency than gelatin sponge andcollagen hemostatic sponge. Composite hemostatic sponge's maximal waterabsorbency could reach 2˜5 times higher than normal gelatin sponge andcollagen hemostatic sponge could. They absorbed water faster and moreefficiently and retained high water absorbency in the fifth and sixth 20s.

Control Experiment 6

Animal Experiment

Objective: To observe the hemostatic effect of modified starchhemostatic sponge in a liver bleeding models.

Experimental Method

An area of 2 cm×1 cm was cut off with a scalpel on the liver surface.Wound depth was 0.3 cm. Hemostatic sponges employed in the experiment tostop bleeding of the wound were: 51# hemostatic sponge B, 66# hemostaticsponge C, composite hemostatic sponge I [51#: medical gelatin (massratio): 1:1], composite hemostatic sponge F [66#: gelatin (mass ratio)1:1], composite hemostatic sponge [66#: carboxymethyl cellulose (massratio): 1:1], and composite hemostatic sponge [66#: collagen (massratio) 10:1]. Simple gelatin sponge and collagen sponge were used ascontrols. When the wound started to bleed, the hemostatic sponges wereimmediately placed on the wound. A medical surgical glove or hemostaticgauze was used to press the wound and to stop the blood stream. 1˜2 minlater, the glove or gauze was released gently to observe the hemostaticeffect and whether the glove or the gauze had adhered to the sponge orthe blot clot and whether re-bleeding occurred as the glove or gauze wasremoved. It was unnecessary to remove modified starch sponge after thebleeding was stopped. The wound was gently irrigated with normal saline.

Results:

All hemostatic sponges containing modified starch in the experimentalgroups had satisfactory hemostatic effect and were convenient to use.Sponges in the experimental groups can absorb moisture/blood immediatelyand form an adhesive sponge-blood coagulation colloid with the blood.Effective control of the bleeding from the liver wound was achieved in1-2 minutes, in comparison to 3-5 min or more with the gelatin spongeand collagen sponge. Upon contact with the blood, hemostatic sponges inthe experimental groups can adhere to the liver wound tissue tightly topromote blood coagulation and seal the bleeding vessels on the woundsurface. The sponges in the experimental groups are elastic and easy touse. They do not adhere to the glove or gauze that are used to press thewound, and do not destroy the blood clot when the glove or gauze isremoved, and therefore do not cause re-bleeding. The gelatin sponge andcollagen hemostatic sponge in the control groups absorbed moisture/bloodslowly, and needed to be pressed for multiple times. They adhered poorlyto the tissue of wound surface, and had poor hemostatic effects.

One skilled in the art will understand that the embodiments of presentinvention, as shown in the drawings and described above are exemplaryonly and not intended to be limited.

It will thus be seen that the objectives of the present invention havebeen fully and effectively achieved. The embodiments have been shown anddescribed for the purposes of illustrating the functional and structuralprinciples of the present invention and are subject to change withoutdeparture from such principles. Therefore, this invention includes allmodifications encompassed within the spirit and scope of the followingclaim.

What is claimed is:
 1. A biocompatible modified starch for effectinghemostasis, anti-adhesion therapy, tissue healing promotion, woundsealing, and wounded tissue bonding by applying directly onto thewounded tissue of the animal, wherein the biocompatible modified starchis a hemostatic powder consisting essentially of biocompatible modifiedstarch particles having water absorbency capacity not lower than 1 timesits own particle weight and a molecular weight 10,000 Daltons or moreand a grain diameter of 10 to 1000 μm, wherein the hemostatic powder hasan adhesiveness sufficient that the hemostatic powder forms astarch-blood coagulation upon contacting the wounded tissue when beingapplied directly onto the wounded tissue of the animal, wherein thebiocompatible modified starch contains one or more groups ofpre-gelatinized starch, acid modified starch, oxidized starch,esterified starch, and cross-linked starch, wherein the biocompatiblemodified starch contains at least pre-gelatinized hydroxypropyl distarchphosphate in pre-gelatinization modification.
 2. A biocompatiblemodified starch for effecting hemostasis, anti-adhesion therapy, tissuehealing promotion, wound sealing, and wounded tissue bonding by applyingdirectly onto the wounded tissue of the animal, wherein thebiocompatible modified starch is a hemostatic powder consistingessentially of biocompatible modified starch particles having waterabsorbency capacity not lower than 1 times its own particle weight and amolecular weight 10,000 Daltons or more and a grain diameter of 10 to1000 μm, wherein the hemostatic powder has an adhesiveness sufficientthat the hemostatic powder forms a starch-blood coagulation uponcontacting the wounded tissue when being applied directly onto thewounded tissue of the animal, wherein the biocompatible modified starchparticles have starch grains with grain diameter 30 to 500 μm, at least95% starch grains in said hemostatic powder with a diameter 30 to 500 μmof in total, wherein the biocompatible modified starch contains one ormore groups of pre-gelatinized starch, acid modified starch, oxidizedstarch, esterified starch, and cross-linked starch, wherein thebiocompatible modified starch contains at least pre-gelatinizedhydroxypropyl distarch phosphate in pre-gelatinization modification. 3.A biocompatible modified starch for effecting hemostasis, anti-adhesiontherapy, tissue healing promotion, wound sealing, and wounded tissuebonding by applying directly onto the wounded tissue of the animal,wherein the biocompatible modified starch is a hemostatic powderconsisting essentially of biocompatible modified starch particles havingwater absorbency capacity not lower than 1 times its own particle weightand a molecular weight 10,000 Daltons or more and a grain diameter of 10to 1000 μm, wherein the hemostatic powder has an adhesiveness sufficientthat the hemostatic powder forms a starch-blood coagulation uponcontacting the wounded tissue when being applied directly onto thewounded tissue of the animal, wherein said biocompatible modified starchcontains one or more groups of pre-gelatinized starch, acid modifiedstarch, oxidized starch, esterified starch, and cross-linked starch,wherein the biocompatible modified starch contains at least onecarboxymethyl starch, hydroxyethyl starch, and cationic starch inpre-gelatinization modification.
 4. A biocompatible modified starch foreffecting hemostasis, anti-adhesion therapy, tissue healing promotion,wound sealing, and wounded tissue bonding by applying directly onto thewounded tissue of the animal, wherein the biocompatible modified starchis a hemostatic powder consisting essentially of biocompatible modifiedstarch particles having water absorbency capacity not lower than 1 timesits own particle weight and a molecular weight 10,000 Daltons or moreand a grain diameter of 10 to 1000 μm, wherein the hemostatic powder hasan adhesiveness sufficient that the hemostatic powder forms astarch-blood coagulation upon contacting the wounded tissue when beingapplied directly onto the wounded tissue of the animal, wherein thebiocompatible modified starch particles have starch grains with graindiameter 30 to 500 μm, at least 95% starch grains in said hemostaticpowder with a diameter 30 to 500 μm of in total, wherein thebiocompatible modified starch contains one or more groups ofpre-gelatinized starch, acid modified starch, oxidized starch,esterified starch, and cross-linked starch, wherein the biocompatiblemodified starch contains at least one carboxymethyl starch, hydroxyethylstarch, and cationic starch in pre-gelatinization modification.